Decreases of metallothionein and aminopeptidase N in renal cancer tissues

Good molecular markers for investigating the biochemical differences between renal cancer and surrounding tissues have not yet been developed. Sixteen kidney samples (clear cell RCC) were investigated to determine the differences in the protein components between renal cancer and surrounding tissues, using HPLC analysis. The metallothionein (MT) and zinc levels were consistently lower in renal cancer tissues compared with in surrounding tissues. The mean concentration of MT in normal tissues surrounding renal tumors was about 15 times higher than that in cancer tissues. An immunohistochemical study confirmed that the expression of MT in renal cancer tissues was lower than that in adjacent normal tissues. The activities of aminopeptidases (APs) were significantly decreased in renal cancer tissues compared with in adjacent normal tissues. An immunohistochemical study and Western blot analysis confirmed that the expression of AP-N in renal cancer tissues was also lower than in adjacent normal tissues. These results suggest that the immunohistochemical detection of MT and AP-N could provide useful information as a pathological diagnostic tool for classifying renal cancer and surrounding tissues.     

CD90在肝细胞肝癌组织中的表达及其与肝癌临床病理特征的关系

目的:探讨CD90在肝细胞肝癌(HCC)患者肿瘤组织中的表达及其与临床病理参数的关系,为HCC的临床治疗提供参考。方法:选择2014年3月-2015年3月在我院接受治疗的HCC患者80例,收集患者肿瘤组织及癌旁组织,另选择同期就诊的80例正常肝脏组织作为对照组。观察并比较CD90在肝癌组织、癌旁组织及正常肝脏组织中的表达情况。结果:CD90在肝癌组织、癌旁组织及正常肝脏组织中呈不同程度阳性表达(P0.05)。CD90在肝癌组织和癌旁组织中的表达显著高于正常肝脏组织,差异具有统计学意义(P0.05)。CD90在肝癌组织中的表达显著高于癌旁组织,差异具有统计学意义(P0.05)。CD90在HCC组织中的表达与肿瘤大小、肝内转移、TMN分期和病理分级有关(P0.05)。结论:CD90在肝癌组织中呈高表达,其表达水平与肝癌临床病理参数有关,可以作为HCC诊断、治疗及预后评估的参考指标。     

Concentrations of some major and minor elements in larynx tissues with and without cancer

In this study, concentrations of some major and minor elements were determined in the larynx tissues with and without cancer, and results obtained were statistically compared. No meaningful differences were found between sodium, potassium, calcium and copper concentrations in cancer tissues, corresponding cancer-free adjacent tissues and in control larynx tissues. Phosphate concentrations of the cancer tissues were higher compared with cancer-free adjacent tissues and control tissues. Iron, zinc and magnesium concentrations were found increased in both cancer and corresponding cancer-free adjacent tissues relative to control values. Intra- and inter-element correlations established within and between groups indicated that relations between elements were also disordered in the cancer tissues. We suggest that the changed element status of cancerous larynx tissues may arise from increased requirements of cancer tissues for some elements such as iron, zinc, magnesium and phosphate.     

Growth and in-vitro metabolism of placental tissues of cows from day 100 to day 250 of gestation.

Weight of placental tissues of cows increased exponentially from Day 100 to Day 250 of gestation, but at much slower relative and absolute rates than fetal weight. In addition, growth rate of fetal placental tissues was less than that of maternal placental tissues. Concentrations of DNA, RNA and protein, however, increased in fetal placental but not in maternal placental tissues. Fetal placental tissues therefore exhibited hyperplasia, which probably contributes to increased functional capacity of the placenta during late gestation. The rate of O2 uptake in vitro was greatest for maternal placental tissues, suggesting that the maternal portion of the placenta accounts for most of the large rate of placental O2 utilization in vivo. Compared with other placental tissues, rate of secretion of macromolecules by intercaruncular endometrium was high, but decreased from Day 100 to 250, suggesting that uterine glandular secretory activity may decrease as gestation advances. Rate of secretion of macromolecules also was high for intercotyledonary tissues and increased with day of gestation, suggesting a role for secretory products of chorioallantois in gravid uterine function.     

Variation in the distribution of trace elements in hepatoma

There are many reports of reduction of zinc level and rise of copper level in serum of patients with liver disease. However, there are a few reports that compare the trace elements in tumor tissues and nontumor tissues of the liver with hepatoma. We studied trace element distribution in tumor tissues and nontumor tissues of liver with hepatoma and compared them with data from normal liver tissues. Zinc (Zn), copper (Cu), selenium (Se), cadmium (Cd), mercury (Hg), and iron (Fe) were chosen as the trace elements to be observed. We observed falls of Zn, Cd, and Hg levels in tumor tissues and the rise of Cu level as a result of this investigation. Zn, Cd, and Hg levels in tumor tissues were significantly lower than those in nontumor tissues and Zn, Cd, and Hg levels in nontumor tissues were significantly lower than in normal liver tissues. This tendency was clearer for Cd and Hg than for Zn. Although the distribution of Cu was not significant, a distribution contrary to that of Zn was shown. These findings indicate that the distribution of Zn, Cd, and Hg can serve as supportive evidence that could be useful as a tumor marker. Selenium showed almost the same accumulation tendency among tumor tissues, nontumor tissues, and normal livers. Although correlation was observed among most metals in the normal liver, there was almost no correlation in tumor tissues.     

Cryopreservation of brain tissue for primary culture.

Factors affecting recovery of brain cells from cryopreserved cerebral tissues of fetal rats were examined based on yields of viable cells on cell culture. Favorable preservation was obtained with freezing small pieces (less than 1 mm cube) of brain tissues rather than whole tissues or dissociated single cells, and use of 10% dimethylsulfoxide as a cryoprotectant in liquid nitrogen. As for cell preparation procedures, cell survival was improved when tissues were heated at 32 degrees C during papain digestion and centrifugation. Under favorable conditions, the number of brain cells recovered from cryopreserved tissues corresponded to 20-30% of those from fresh control tissues. Immunocytochemical characteristics of cultured neurons, astrocytes, and oligodendrocytes from cryopreserved and fresh tissues were indistinguishable. Semi-quantitive analyses of microtubule-associated protein-2 (MAP-2) and synaptophysin revealed that there was no difference in the amounts of these markers between cultures from both fresh and cryopreserved tissues. These results suggest that most of all cell types including neurons were equally susceptible to the cryopreservation procedures. We concluded that cryopreservation in liquid nitrogen is an effective method for preservation of embryonic brain tissues for later use in cell culture studies.     

Tissue specific DNA methylation of CpG islands in normal human adult somatic tissues distinguishes neural from non-neural tissues

Although most CpG islands are generally thought to remain unmethylated in all adult somatic tissues, recent genome-wide approaches have found that some CpG islands have distinct methylation patterns in various tissues, with most differences being seen between germ cells and somatic tissues. Few studies have addressed this among human somatic tissues and fewer still have studied the same sets of tissues from multiple individuals. In the current study, we used Restriction Landmark Genomic Scanning to study tissue specific methylation patterns in a set of twelve human tissues collected from multiple individuals. We identified 34 differentially methylated CpG islands among these tissues, many of which showed consistent patterns in multiple individuals. Of particular interest were striking differences in CpG island methylation, not only among brain regions, but also between white and grey matter of the same region. These findings were confirmed for selected loci by quantitative bisulfite sequencing. Cluster analysis of the RLGS data indicated that several tissues clustered together, but the strongest clustering was in brain. Tissues from different brain regions clustered together, and, as a group, brain tissues were distinct from either mesoderm or endoderm derived tissues which demonstrated limited clustering. These data demonstrate consistent tissue specific methylation for certain CpG islands, with clear differences between white and grey matter of the brain. Furthermore, there was an overall pattern of tissue specifically methylated CpG islands that distinguished neural tissues from non-neural.     

The value of frozen cartilage tissues without cryoprotection for genetic conservation

Animal tissues frozen without cryoprotection are thought to be inappropriate for use as a donor for somatic cell nuclear transfer (SCNT) studies. Cells in tissues that have been frozen without a cryoprotectant are commonly thought to be dead or to have lost genomic integrity. However, in this study we show that the frozen auricular cartilage tissues of anatolian buffalo contain a considerable number of viable healthy cells. The cells in auricular cartilage tissues are resistant to cryo-injury at −80 °C. Primary cell cultures were established from defrosted ear tissues which were frozen without cryoprotectant. The growth and functional characteristics of primary cell cultures are characterized according to cell growth curve, cell cycle analysis, karyotype and GAG synthesis. The results indicate that frozen cartilage tissues could be valuable materials for the conservation of species and SCNT technology.     

LR8 expression in fibroblasts of healthy and fibrotic human tissues

LR8 gene was first reported in a subpopulation of cultured human lung fibroblasts expressing the receptor for C1q-globular domain, and it was not detectable in cultured endothelial cells and smooth muscle cells. LR8 mRNA levels were higher in fibrotic lungs. In this study we assessed LR8 production in human tissues and determined if the distribution of fibroblasts producing LR8 is affected in fibrosis. Normal and fibrotic tissue sections from human liver, lung and kidneys were immunostained with antibodies to LR8 and examined for the presence of fibroblasts staining positively and negatively. The cells were also examined for co-expression of α-smooth muscle actin (SMA), a marker for myofibroblasts. The results showed that LR8 was expressed by fibroblasts, smooth muscle cells, endothelial cells, bile duct cells, pulmonary alveolar cells and distal and proximal kidney tubule cells. Connective tissues of normal and fibrotic tissues contained fibroblasts staining positively and negatively with anti- LR8 antibody. The number of LR8-positive cells was higher in fibrotic tissues, but differences were not statistically significant. Fibroblasts producing both LR8 and SMA were present in higher numbers in fibrotic tissues as compared to normal tissues and the differences were statistically significant (p<0.05). Our results show that fibroblast subtypes differing in LR8 expression are present in human tissues, and that in fibrotic tissues cells co-expressing LR8 and SMA are present. Our results indicate that LR8 expressing cells may participate in the early stages of fibrotic diseases and that fibroblasts expressing LR8, not LR8 negative cells, have potential to become myofibroblasts in fibrotic tissues.     

组织的前向散射、后向漫射

人体组织的光学性质是光医学和组织光学领域中的最基本的研究课题之一。当激光与生物肌体相互作用后所产生的各种生物效应,这些即取决于所施加的激光参量,又决定于被作用组织的结构、物理和生物特性。从激光诊断或治疗的角度考虑,由于光或激光只有透过组织,才能到达组织的深部和内部器官。所以,首要解决的问题是组织对光的通透性及其机理的研究。光通过组织时,光强和光的偏振状态会发生变化。令He－Ne激光照射组织,分别在组织的透射方和反射方探测光强．当光垂直（θ＝0°）或以角度θ入射到具有一定厚度的组织时,探测器以α的角度测量前后向的光强分布．根据实验数据分别绘制了前向散射和后向漫射光强的极坐标曲线．当θ－0°、探测器的角度为α＝0°时光的强度最大（前向散射）．旋转探测器的角度（改变α）探测到的光强其图形为一个圆。当入射光的角度θ由0°变到60°时,光的强度降低了31．65％．此时旋转探测器（改变α）探测到的光强分布图形变为椭圆,并且和原点相切,和θ＝0°的图形相比,此时图形的半宽度增加。当入射光的角度θ固定为0°、探测器角度α变化接近180°时光的后向漫射强度最大,其图形为一个圆。比较前向散射和后向漫射的光强分布图可看出光强分布基本上对称。如将两幅     

SARS-CoV N蛋白与Smad3相互作用并调节TGF-β信号通路的形态学初步研究

目的应用病理学实验技术,初步描述SARS-CoV N蛋白与TGF-β信号通路中Smad3蛋白之间相互作用的情况,及其对TGF-β信号通路下游转录产物生成的影响。方法对比观察SARS-CoV感染的恒河猴肺组织及正常猴肺组织的形态学特征,应用免疫组织化学染色法,观察猴肺组织中转化生长因子-β（transforming growth factor-β,TGF-β）及1型纤维蛋白溶酶原活化抑制剂（plasminogen activator inhibitor-1,PAI-1）的肺组织表达;SARS-CoVNucleocapsid蛋白与Smad3双染观察二者在肺组织中的共定位情况;应用Masson染色法比较感染与正常猴肺组织中胶原含量的差异,结合分子生物学检测肺组织中TGF-β、PAI-1及Ⅰ型胶原α2链（α2 chain of typeⅠcollagen,COLlA2）的表达,探讨N蛋白影响TGF-β信号通路的影响。结果PCR结果显示TGF-β、PAI-1及COL1A2在SARS-CoV感染的猴肺组织中的表达量较正常猴肺有明显增高,免疫组织化学染色结果提示,在病毒感染的猴肺组织内TGF-β、PAI-1染色呈现强阳性,同时SARS-CoV Nucleocapsid蛋白与Smad3双染结果显示,二者在感染的猴肺组织中存在明显的共定位现象。Masson染色结果提示,病毒感染的猴肺组织胶原含量较正常猴肺组织显著增加。结论我们的研究结果为说明SARS-CoV Nucleocapsid蛋白参与调节TGF-β信号通路,促进肺组织纤维化提供了有力的形态学实验证明,为进一步理解SARS-CoV感染引起肺组织纤维化的机制提供了的较为明确体内实验数据。     

Complex carbohydrates in cartilaginous and other tissues of cartilage matrix deficiency (cmd/cmd) mice as studied by light microscopic histochemical methods

Summary Mice homozygous for the autosomal recessive gene, cartilage matrix deficiency (cmd/cmd) are characterized by disproportionate dwarfism and cleft palate and are suffered from genetic failure in biosynthesis of cartilagetype proteoglycans. To investigate histochemical aspects of the defects in cartilaginous and other tissues of the mutant mice, complex carbohydrates in the tissues have been studied by methods of light microscopy. As a result of the present investigations, the intercellular matrix of cartilaginous tissues from the mutant was shown to exhibit weaker positive reactions for sulfated and acidic complex carbohydrates, as compared with the corresponding tissues from the control. In certain muscular and nervous tissues from the mutant, the reactions for sulfated complex carbohydrates tended to be weaker in intensity than those in the control. Furthermore, the dermal tissues of the mutant showed apparent mastocytosis. All these results indicate that there exist accessory effects in non-cartilaginous tissues in addition to the defects of cartilaginous tissues in the mutant mice.     

Genome-wide profiling of gene expression in 29 normal human tissues with a cDNA microarray.

We have performed a comprehensive analysis of the expression profiles in 25 adult and 4 fetal human tissues by means of a cDNA microarray consisting of 23,040 human genes. This study revealed a number of genes that were expressed specifically in each of those tissues. Among the 29 tissues examined, 4,080 genes were highly expressed (at least a five-fold expression ratio) in one or only a few tissues and 1,163 of those were expressed exclusively (more than a ten-fold higher expression ratio) in a particular tissue. Expression of some of the genes in the latter category was confirmed by northern analysis. A hierarchical clustering analysis of gene-expression profiles in nerve tissues (adult brain, fetal brain, and spinal cord), lymphoid tissues (bone marrow, thymus, spleen, and lymph node), muscle tissues (heart and skeletal muscle), or adipose tissues (mesenteric adipose and mammary gland) identified a set of genes that were commonly expressed among related tissues. These data should provide useful information for medical research, especially for efforts to identify tissue-specific molecules as potential targets of novel drugs to treat human diseases.     

Relationship between genetic variation in thermal stability and electrophoretic mobility of mouse β-galactosidase

We have examined the relationships between the genetic determinants for mouse beta-galactosidase heat stability and electrophoretic mobility, in order to clarify previous reports indicating that a variation for enzyme heat stability is restricted to kidney while that for electrophoretic mobility is expressed in all tissues. We find that the two phenotypes show concordant strain distributions and cosegregate in genetic crosses. In contrast to a previous report, the thermal stability variation is expressed in all tissues, although the absolute rate of enzyme inactivation is tissue specific. The evidence supports the notion that a single beta-galactosidase structural locus is expressed in all tissues and that the differences in enzyme stability between tissues results from posttranslational enzyme modification.     

Fetal Tissue Banking for Transplantation: Characteristics of the Donor Population and Considerations for Donor and Tissue Screening

We initiated this study to evaluate the suitability for therapeutic use in transplantation of tissues obtained from human abortuses. We have developed protocols for the collection, handling and preservation of hepatic stem cells from electively aborted embryos and have developed methods for assessment of the cells so derived and processed. In this paper we present our findings regarding screening of potential donors, acquisition of fetal tissues, and assessment of the tissues for potentially infectious contaminants. We assess the suitability of the tissue donors according to current standards used for donors of commonly transplanted tissues (e.g., bone grafts, skin grafts and heart valves) and present data regarding the real availability of tissues from elective abortion procedures that would meet those standard tissue banking criteria.We specifically evaluated the donor's willingness to provide a blood sample for testing, conducted a detailed interview similar to those used for typical organ and tissue donors, and assessed the type and incidence of contamination in collected tissues. We find that although many women are willing to consent to use of the tissues for transplantation, attrition from the study for various reasons results in few fetal organs ultimately realistically available for transplantation. Typical reasons for attrition include: unwillingness to have a blood sample drawn or tested, positive serology results, social/medical high risk factors for acquisition of transmissible disease, no identifiable organs available, and unacceptable microbial contamination. Thus, although it might seem that due to the numbers of abortions performed annually, that there would be substantial numbers of suitable tissues available, only a small proportion are truly suitable for transplantation.     

RNA干扰肝细胞肝癌中PECAM-1的表达对肿瘤细胞侵袭及增殖的影响

目的:探讨PECAM-1在肝细胞肝癌(Hepatocellular carcinoma,HCC)组织中的表达及意义。方法:选择2013年5月-2015年6月在我院接受治疗的HCC患者100例,收集肝癌患者HCC组织及癌旁组织,另选取100例正常肝脏组织作为对照组。应用免疫组织化学法检测PECAM-1在肝癌组织、癌旁组织以及正常肝脏组织中的阳性表达。利用小分子干扰RNA技术(si RNA)构建低表达的PECAM-1,并转染至肝癌细胞中抑制PECAM-1的表达。应用Transwell小室法检测肝癌细胞的侵袭能力,CCK-8法检测肝癌细胞的增殖能力。结果:PECAM-1在肝癌组织、癌旁组织及正常肝脏组织中呈不同程度阳性表达(P0.05);PECAM-1在肝癌组织及癌旁组织中的表达显著高于正常肝脏组织,差异具有统计学意义(P0.05);PECAM-1在肝癌组织中的表达显著高于癌旁组织,差异具有统计学意义(P0.05);转染si RNA PECAM-1后,肝癌细胞中PECAM-1 m RNA的表达水平明显下降,PECAM-1蛋白表达也明显降低,差异具有统计学意义(P0.05);转染si RNA PECAM-1后,肝癌细胞侵袭及增殖能力明显降低,差异具有统计学意义(P0.001)。结论:PECAM-1在肝癌患者血清中高表达,PECAM-1 si RNA能够抑制肝癌细胞的侵袭及增殖能力,提示PECAM-1可作为预测肝癌发生及发展的临床指标。     

Transcriptomic characterization of gall tissue of Japanese elm tree (Ulmus davidiana var. japonica) induced by the aphid Tetraneura nigriabdominalis

Insect galls are abnormal plant tissues induced by parasitic insect(s) for use as their habitat. In previous work, we suggested that gall tissues induced by the aphid Tetraneura nigriabdominalis on Japanese elm trees are less responsive than leaf tissues to jasmonic acid (JA), which is involved in the production of volatile organic compounds as a typical defensive reaction of plants against attack by insect pests. A comprehensive analysis of gene expression by RNA sequencing indicated that the number of JA responsive genes was markedly lower in gall tissues than in leaf tissues. This suggests that gall tissues are mostly defective in JA signaling, although JA signaling is not entirely compromised in gall tissue. Gene ontology analysis sheds light on some stress-related unigenes with higher expression levels in gall tissues, suggesting that host plants sense aphids as a biotic stress but are defective in the JA-mediated defense response in gall tissues.     

Effect of castration monotherapy on the levels of adrenal androgens in cancerous prostatic tissues

The mechanism accounting for the development of castration-resistant prostate cancer (CRPC) remains unclear. Studies in CRPC tissues suggest that, after androgen deprivation therapy (ADT), the adrenal androgens may be an important source of testosterone (T) and 5-alpha dihydrotestosterone (DHT) in CRPC tissues. To clarify the role of adrenal androgens in the prostatic tissues (prostatic tissue adrenal androgens) during ADT, we developed a high sensitive and specific quantification method for the levels of androgens in prostatic tissue using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Human prostatic tissues were purified using mixed-mode reversed-phase, strong anion exchange Oasis cartridges (Oasis MAX). Analysis of steroids was performed using LC-MS/MS after picolinic acid derivatization. The validation tests showed that our method of quantitative analysis was precise and sensitive enough for the quantification of dehydroepiandrosterone (DHEA), androstenedione, androstenediol, T, and DHT in the prostatic tissue. The levels of adrenal androgens in prostate cancer tissues after ADT were similar to those in untreated PCa. Especially, DHEA was the most existing androgen precursor in PCa tissues after ADT. The levels of DHEA were high in PCa tissues, irrespective of ADT. We assumed that DHEA played a significant role in the synthesis of T and DHT in PCa tissues after ADT.